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EGFR STAR ELISA Assay kit
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Description:
EGFR STAR ELISA Kit
Trade Name:
Upstate (Millipore)
Qty/Pk:
96 assays
Background Information:
I. TEST PRINCIPLE
The UPSTATE® colorimetric STAR (Signal Transduction Assay Reaction) ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to detect specific levels of signaling targets in whole cell extracts. The EGFR plate is coated with a specific mouse monoclonal anti-EGFR capture antibody. Sample lysate or the standard included in the kit are incubated in the microwells allowing EGFR antigen to be captured in the plate wells. The plate is then washed to remove any un-bound non-nspecific material. A specific rabbit anti-EGFR antibody is added to detect the captured full length EGFR on the plate well. The unbound detection antibody is washed away followed by incubation with an HRP-conjugated anti-rabbit antibody. After the addition of TMB substrate and stop solution the absorbance is measured at 450 nm using a plate reader. This allows for a sensitive enzymatic detection of the sample.
The entire assay takes less than 5 hours to complete with minimal hands-on time. Many of the reagents are supplied in ready-to use formulations for ease of use. The kit also includes a standard that is run as both a positive control and to to generate a standard curve for EGFR measurement.
II. EGFR BACKGROUND
Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that plays a key role in the regulation of essential normal cellular processes and in the pathophysiology of hyperproliferative diseases such as cancer. Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase known to be essential for mediation of both proliferative and survival signals to cells. Activation of the EGFR signaling pathway has been linked with increased cell proliferation, angiogenesis, metastasis and decreased apoptosis. EGFR is found in most solid tumors and its autophosphorylation activates the intrinsic kinase activity toward heterologous substrates, as well as creating docking sites for adapter proteins for multiple adapter proteins to bind and nucleate signaling complexes that activate the Ras, PI3 Kinase, and PLC pathways.
Many of the EGFR mutations are directly related to various diseases and are the target of numerous drugs. For instance, the amino acid substitution L858R is one of several heterozygous mutations that have been identified in Non-Small-Cell Lung Cancer (NSCLC) patients who have clinical responses to the EGFR inhibitor Iressa®. There is some evidence that these mutations result in elevated activity and enhanced sensitivity to Iressa®. In patients with tumors bearing Iressa®-sensitive mutations, resistant subclones containing an additional EGFR mutation, T790M, emerge in the presence of the drug. It has been shown experimentally that the T790M mutation leads to high-level functional resistance to Iressa. In patients with tumors bearing Iressa®-sensitive mutations (eg. L858R, L861Q), resistant subclones containing the T790M mutation emerge in the presence of the drug.
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Key Applications:
ELISA
Species Reactivity:
Human
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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UniProt Summary:
FUNCTION: SwissProt: P00533 # Isoform 2/truncated isoform may act as an antagonist.
SIZE: 1210 amino acids; 134277 Da
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